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Microsatellite triple ternary PCR detection method of marsupenaeus japonicas

A detection method and microsatellite technology, which are applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., to improve detection efficiency, reduce electrophoresis detection time and cost, and save PCR and electrophoresis reagents.

Inactive Publication Date: 2015-12-23
TIANJIN UNIV OF COMMERCE
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  • Claims
  • Application Information

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Problems solved by technology

There is no relevant report on multiple PCR detection technology in Penaeus japonicus

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  • Microsatellite triple ternary PCR detection method of marsupenaeus japonicas
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  • Microsatellite triple ternary PCR detection method of marsupenaeus japonicas

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Embodiment 1

[0020] (1), primer design and combination

[0021] Use TandemRepeatFinder to find microsatellite sequences from random clone sequencing sequences, select complete flanking sequences on both sides of the core repeat sequence and use PremierPrimer to design primers. MPprimer evaluates the combination of primers, and takes the absolute value of △G to be less than 3, which reduces the probability of secondary structure formation and improves the specificity and efficiency of PCR reactions. Through the combination of many pairs of microsatellite primers, the multiple PCR system combination, PCR reaction conditions and reaction procedures of Penaeus japonicus are optimized, and finally a kind of Microsatellite 3-fold PCR reaction system and reaction of Penaeus japonicus according to the present invention are established. Procedures and detection methods. 3 pairs of P. japonicus primers selected by the present invention are:

[0022] SEQ036 forward primer sequence: 5'-AAGGGAATTTGAG...

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Abstract

The invention discloses a microsatellite triple ternary PCR detection method of marsupenaeus japonicas. The method includes the steps that three pairs of PCR primers are designed and synthesized; the total DNA is extracted; combination, a system and a reaction program of the ternary PCR reactions are provided; PCR products are detected. The efficiency of the microsatellite triple ternary PCR detection method of marsupenaeus japonicas can be tripled compared with an original method. The method has the advantages that the cost and time are saved, and the method is efficient, economical, high in repeatability, simple, feasible, beneficial to detection and the like and can be applied and popularized with respect to breeding management, family identification, individual recognition, genetic relationship identification, improved breed breeding, linkage map construction, genetic diversity detection and the like of marsupenaeus japonicas.

Description

technical field [0001] The invention belongs to molecular marker auxiliary technology of genetic breeding of molecular biology, in particular to a microsatellite triple PCR detection method of Penaeus japonicus. Background technique [0002] Penaeus japonicus is widely distributed in the Indian Ocean, the Western Pacific, and the coasts of Japan and the southeast coast of my country. It is an important breeding object in my country's coastal areas. The use of molecular markers is one of the important ways to select and breed improved Penaeus japonicus. At present, microsatellites are widely used molecular markers. Microsatellites are widely distributed in the genome, with high polymorphism, good stability, and co-dominant inheritance. It is widely used in pedigree identification, individual identification, gene linkage analysis, genetic linkage map construction, genetic diversity detection and many other fields. Conventional microsatellite detection technology is single-pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2525/151
Inventor 董世瑞栾生王素英王博玮陈瑾
Owner TIANJIN UNIV OF COMMERCE
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